{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,17]],"date-time":"2026-01-17T16:26:18Z","timestamp":1768667178404,"version":"3.49.0"},"reference-count":45,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T00:00:00Z","timestamp":1768435200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"Natural Science Foundation of China","doi-asserted-by":"crossref","award":["32260473"],"award-info":[{"award-number":["32260473"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001809","name":"Natural Science Foundation of China","doi-asserted-by":"crossref","award":["32360511"],"award-info":[{"award-number":["32360511"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100001809","name":"Natural Science Foundation of China","doi-asserted-by":"crossref","award":["32460510"],"award-info":[{"award-number":["32460510"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Biological Breeding Project of Gansu Academy of Agricultural Sciences","award":["2026GAAS01"],"award-info":[{"award-number":["2026GAAS01"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Plants"],"abstract":"<jats:p>Leaf color mutants are commonly characterized by altered chlorophyll content and aberrant chloroplast development, making them valuable models for investigating photosynthetic mechanisms and chloroplast biogenesis. In this study, an albino mutant was isolated from a population of transgenic maize breeding lines. Genetic analysis indicated that the mutant phenotype is inherited in a Mendelian manner and is controlled by a single nuclear locus. This was supported by a \u03c72 test performed on the T2 generation, which confirmed a segregation ratio consistent with 3:1 (176:68, \u03c72 = 1.07 &lt; \u03c720.05 = 3.84, p &gt; 0.05). Microscopic examination revealed the absence of normally developed chloroplasts in mutant cells. Further expression analysis of chloroplast genes via Northern blotting and quantitative real-time PCR (qRT-PCR) suggested that the mutation impairs the regulation of plastid-encoded polymerase (PEP)-dependent chloroplast gene expression. Notably, PCR-based co-segregation analysis indicated that the mutant phenotype is associated with the entire inserted vector sequence, rather than a point mutation or a small genomic deletion. In conclusion, this paper reports the isolation and phenotypic characterization of an etiolated mutant from a transgenic maize breeding population, including comparative ultrastructural analysis of chloroplasts, co-segregation validation, and chloroplast gene expression profiling. These results enhance our understanding of the physiological and molecular mechanisms underlying chlorophyll-deficient mutations in plants.<\/jats:p>","DOI":"10.3390\/plants15020266","type":"journal-article","created":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T11:13:21Z","timestamp":1768475601000},"page":"266","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Identification of a Chlorophyll-Deficient Mutant in Maize Associated with Exogenous Vector Insertion"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1773-6175","authenticated-orcid":false,"given":"Wenqi","family":"Zhou","sequence":"first","affiliation":[{"name":"Maize Research Center of Gansu Province, Crop Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haoyue","family":"Wang","sequence":"additional","affiliation":[{"name":"Maize Research Center of Gansu Province, Crop Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chunxia","family":"Liang","sequence":"additional","affiliation":[{"name":"Maize Research Center of Gansu Province, Crop Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Haijun","family":"He","sequence":"additional","affiliation":[{"name":"Maize Research Center of Gansu Province, Crop Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yongsheng","family":"Li","sequence":"additional","affiliation":[{"name":"Maize Research Center of Gansu Province, Crop Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaorong","family":"Lian","sequence":"additional","affiliation":[{"name":"Maize Research Center of Gansu Province, Crop Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaojuan","family":"Wang","sequence":"additional","affiliation":[{"name":"Maize Research Center of Gansu Province, Crop Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaoyun","family":"Dong","sequence":"additional","affiliation":[{"name":"Maize Research Center of Gansu Province, Crop Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zengke","family":"Ma","sequence":"additional","affiliation":[{"name":"Maize Research Center of Gansu Province, Crop Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhongxiang","family":"Liu","sequence":"additional","affiliation":[{"name":"Maize Research Center of Gansu Province, Crop Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuqian","family":"Zhou","sequence":"additional","affiliation":[{"name":"Maize Research Center of Gansu Province, Crop Research Institute, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2026,1,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"110743","DOI":"10.1016\/j.scienta.2021.110743","article-title":"Insights into the Molecular Basis of a Yellow Leaf Color Mutant (ym) in Tomato (Solanum lycopersicum)","volume":"293","author":"Cheng","year":"2022","journal-title":"Sci. Hortic."},{"key":"ref_2","first-page":"319","article-title":"Fine Mapping of a Yellowing Mutant Gene Zmet9 in Maize","volume":"26","author":"Li","year":"2025","journal-title":"J. Plant Genet. Resour."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1233","DOI":"10.1093\/mp\/ssp093","article-title":"Blocking the Metabolism of Starch Breakdown Products in Arabidopsis Leaves Triggers Chloroplast Degradation","volume":"2","author":"Stettler","year":"2009","journal-title":"Mol. Plant"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1105\/tpc.014787","article-title":"Maize Mutant Lacking Chloroplast FtsY Exhibits Pleiotropic Defects in the Biogenesis of Thylakoid Membranes","volume":"16","author":"Asakura","year":"2004","journal-title":"Plant Cell"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"342","DOI":"10.1104\/pp.110.161414","article-title":"A Mutation in Arabidopsis Seedling Plastid Development1 Affects Plastid Differentiation in Embryo-Derived Tissues during Seedling Growth","volume":"155","author":"Ruppel","year":"2011","journal-title":"Plant Physiol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1016\/j.tplants.2004.04.001","article-title":"Genetics and Genomics of Chloroplast Biogenesis: Maize as a Model System","volume":"9","author":"Stern","year":"2004","journal-title":"Trends Plant Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"37098","DOI":"10.1074\/jbc.M504805200","article-title":"Inactivation of Organellar Glutamyl- and Seryl-tRNA Synthetases Leads to Developmental Arrest of Chloroplasts and Mitochondria in Higher Plants","volume":"280","author":"Kim","year":"2005","journal-title":"J. Biol. Chem."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1046\/j.1365-313x.2000.00672.x","article-title":"Cloning and Functional Expression of the Gene Encoding the Key Enzyme for Chlorophyll b Biosynthesis (CAO) from Arabidopsis thaliana","volume":"21","author":"Oster","year":"2000","journal-title":"Plant J."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"805","DOI":"10.1007\/s11103-005-2066-9","article-title":"Differential Regulation of Chlorophyll a Oxygenase Genes in Rice","volume":"57","author":"Lee","year":"2005","journal-title":"Plant Mol. Biol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1362","DOI":"10.1105\/tpc.106.042911","article-title":"Rice NON-YELLOW COLORING 1 is Involved in Light-Harvesting Complex II and Grana Degradation during Leaf Senescence","volume":"19","author":"Kusaba","year":"2007","journal-title":"Plant Cell"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1011","DOI":"10.1111\/j.1365-313X.2009.03930.x","article-title":"AtECB2, a Pentatricopeptide Repeat Protein, Is Required for Chloroplast Transcript accD RNA Editing and Early Chloroplast Biogenesis in Arabidopsis thaliana","volume":"59","author":"Yu","year":"2009","journal-title":"Plant J."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1542","DOI":"10.1073\/pnas.1221738110","article-title":"Unanticipated Regulatory Roles for Arabidopsis Phytochromes Revealed by Null Mutant Analysis","volume":"110","author":"Hu","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_13","unstructured":"Xue, Y.J. (2023). Mapping and Functional Analysis of Genes Related to Photosynthesis in Maize. [Ph.D. Thesis, Jilin University]."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2215","DOI":"10.1271\/bbb.68.2215","article-title":"Roles of Chloroplast RNA Polymerase Sigma Factors in Chloroplast Development and Stress Response in Higher Plants","volume":"68","author":"Kanamaru","year":"2004","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"e0110","DOI":"10.1199\/tab.0110","article-title":"Chloroplast Biogenesis: Control of Plastid Development, Protein Import, Division and Inheritance","volume":"6","author":"Sakamoto","year":"2008","journal-title":"Arab. Book"},{"key":"ref_16","first-page":"14","article-title":"Advances in the Study of Leaf Color Mutants in Maize","volume":"13","author":"Li","year":"2019","journal-title":"South. Agric."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1038\/nrg1271","article-title":"Endosymbiotic Gene Transfer: Organelle Genomes Forge Eukaryotic Chromosomes","volume":"5","author":"Timmis","year":"2004","journal-title":"Nat. Rev. Genet."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"809","DOI":"10.1126\/science.277.5327.809","article-title":"Mitochondrial and Chloroplast Phage-Type RNA Polymerases in Arabidopsis","volume":"277","author":"Hedtke","year":"1997","journal-title":"Science"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/S0074-7696(08)62145-2","article-title":"Organellar RNA Polymerases of Higher Plants","volume":"190","author":"Hess","year":"1999","journal-title":"Int. Rev. Cytol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Bock, R. (2007). Transcription and Transcriptional Regulation in Plastids. Cell and Molecular Biology of Plastids, Springer.","DOI":"10.1007\/978-3-540-75376-6"},{"key":"ref_21","first-page":"73","article-title":"Evolutionary analysis of chloroplast RNA editing in terrestrial plants","volume":"1","author":"Wan","year":"2013","journal-title":"Biotechnol. Bull."},{"key":"ref_22","unstructured":"Huang, W.F. (2020). Functional Identification and Mechanism Analysis of OsPPR16 during Early Chloroplast Development in Rice. [Ph.D. Thesis, Huazhong Agricultural University]."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1433","DOI":"10.1111\/nph.19243","article-title":"Interaction of PALE CRESS with PAP2\/pTAC2 and PAP3\/pTAC10 Affects the Accumulation of Plastid-Encoded RNA Polymerase Complexes in Arabidopsis","volume":"240","author":"Wang","year":"2023","journal-title":"New Phytol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1127","DOI":"10.1016\/j.cell.2024.01.026","article-title":"Cryo-EM Structures of the Plant Plastid-encoded RNA Polymerase","volume":"187","author":"Wu","year":"2024","journal-title":"Cell"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1016\/j.jplph.2012.01.019","article-title":"Evaluation of Candidate Reference Genes for qPCR in Maize","volume":"169","author":"Manoli","year":"2012","journal-title":"Plant Physiol."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Lv, Z., Zhao, W., Kong, S., Li, L., and Lin, S. (2023). Overview of Molecular Mechanisms of Plant Leaf Development: A Systematic Review. Front. Plant Sci., 14.","DOI":"10.3389\/fpls.2023.1293424"},{"key":"ref_27","unstructured":"Li, R.N. (2024). Identification of PfkB Type Carbohydrate Kinase Alleles in Maize. [Ph.D. Thesis, Huazhong Agricultural University]."},{"key":"ref_28","unstructured":"Cheng, Y.T. (2024). Molecular Mechanism of Chloroplast SUFB on SUFBC2D Complex. [Ph.D. Thesis, Yangzhou University]."},{"key":"ref_29","first-page":"2093","article-title":"Chloroplast Genome Sequencing and Transcriptome Sequencing Provide Insights into the Developmental Mechanism of the Variegated Mutant of Clivia miniata","volume":"41","author":"Li","year":"2022","journal-title":"Plant Cell Rep."},{"key":"ref_30","unstructured":"Yan, X.Q. (2021). Identification and Gene Mapping of a Green-Turning Mutant wsl887 in Rice. [Ph.D. Thesis, Guizhou University]."},{"key":"ref_31","unstructured":"Xing, A.M. (2020). Functional Analysis of the Chaperone Protein Gene PDM1 in Arabidopsis thaliana. [Ph.D. Thesis, Nanjing Agricultural University]."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"460","DOI":"10.1111\/j.0960-7412.2004.02060.x","article-title":"Chloroplast Development Affects Expression of Phage-Type RNA Polymerases in Barley Leaves","volume":"38","author":"Emanuel","year":"2004","journal-title":"Plant J."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"171","DOI":"10.1046\/j.1365-313X.2002.01349.x","article-title":"Comparative Analysis of Plastid Transcription Profiles of Entire Plastid Chromosomes from Tobacco Attributed to Wild-Type and PEP-Deficient Transcription Machineries","volume":"31","author":"Legen","year":"2002","journal-title":"Plant J."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"5152","DOI":"10.1093\/nar\/gkn492","article-title":"A Member of the Whirly Family is a Multifunctional RNA- and DNA-Binding Protein That is Essential for Chloroplast Biogenesis","volume":"36","author":"Prikryl","year":"2008","journal-title":"Nucleic Acids Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1105\/tpc.111.089441","article-title":"The Primary Transcriptome of Barley Chloroplasts: Numerous Noncoding RNAs and the Dominating Role of the Plastid-Encoded RNA Polymerase","volume":"24","author":"Zhelyazkova","year":"2012","journal-title":"Plant Cell"},{"key":"ref_36","first-page":"1233","article-title":"A Natural Variation in PLEIOTROPIC DEVELOPMENTAL DEFECTS Uncovers a Crucial Role for Chloroplast tRNA Modification in Translation and Plant Development","volume":"32","author":"Lu","year":"2020","journal-title":"Plant Cell"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1016\/S0022-2836(61)80072-7","article-title":"Genetic Regulatory Mechanisms in the Synthesis of Proteins","volume":"3","author":"Jacob","year":"1961","journal-title":"J. Mol. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3120","DOI":"10.3724\/SP.J.1006.2022.12077","article-title":"Genetic Analysis and Gene Mapping of Leaf Vein Yellowing Mutant yml in Rice","volume":"48","author":"Guo","year":"2022","journal-title":"Acta Agron. Sin."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1038\/nbt.2095","article-title":"Genome Sequencing Reveals Agronomically Important Loci in Rice Using MutMap","volume":"30","author":"Abe","year":"2012","journal-title":"Nat. Biotechnol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1038\/nbt.3188","article-title":"MutMap Accelerates Breeding of a Salt-Tolerant Rice Cultivar","volume":"33","author":"Takagi","year":"2015","journal-title":"Nat. Biotechnol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1111\/tpj.12105","article-title":"Rapid Mapping of Quantitative Trait Loci in Rice by Whole Genome Resequencing","volume":"74","author":"Chi","year":"2013","journal-title":"Plant J."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"573","DOI":"10.1104\/pp.108.116194","article-title":"The Pentratricopeptide Repeat Protein DELAYED GREENING1 Is Involved in the Regulation of Early Chloroplast Development and Chloroplast Gene Expression in Arabidopsis","volume":"147","author":"Chi","year":"2008","journal-title":"Plant Physiol."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1006\/meth.2001.1262","article-title":"Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2\u2212\u0394\u0394CT Method","volume":"25","author":"Livak","year":"2001","journal-title":"Methods"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1420","DOI":"10.1104\/pp.112.204198","article-title":"Effects of Reduced Chloroplast Gene Copy Number on Chloroplast Gene Expression in Maize","volume":"160","author":"Udy","year":"2012","journal-title":"Plant Physiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1487","DOI":"10.1111\/jipb.13890","article-title":"DSD1\/ZmICEb Regulates Stomatal Development and Drought Tolerance in Maize","volume":"67","author":"Zhou","year":"2025","journal-title":"J. Integr. Plant Biol."}],"container-title":["Plants"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2223-7747\/15\/2\/266\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,1,17]],"date-time":"2026-01-17T05:53:36Z","timestamp":1768629216000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2223-7747\/15\/2\/266"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,1,15]]},"references-count":45,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2026,1]]}},"alternative-id":["plants15020266"],"URL":"https:\/\/doi.org\/10.3390\/plants15020266","relation":{},"ISSN":["2223-7747"],"issn-type":[{"value":"2223-7747","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,1,15]]}}}